摘要
目的研究瞬时通道蛋白V1(transient receptor potential vanilloid 1,TRPV1,又称辣椒素受体)、瞬时通道蛋白M8(transient receptor potential melastatin member 8,TRPM8)在葡聚糖硫酸钠(dextran sulfate sodiμm,DSS)诱导下的实验性结肠炎小鼠肠道和脊髓背根神经节(dorsal root ganglion,DRG)中的表达,进一步探讨两者间的联系及相关通路。方法选取C57BL/6雄性小鼠20只,随机分为空白对照组和DSS组,每组各10只。DSS组使用质量浓度为30g/L的DSS共7天构建结肠炎模型,每天同一时刻观察记录小鼠的毛发、体质量、粪便性状(颗粒状、稀便、血便等)、肛周情况(红肿、便血等),给予疾病活动指数(disease activity index,DAI)评分,根据病理切片进行组织病理学评分,Western blot法检测结肠组织TRPV1、TRPM8、Gαq蛋白的表达,实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测结肠组织白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)的mRNA表达水平,免疫荧光检测结肠组织和DRG中TRPV1、TRPM8及Gαq的定位及表达。结果与空白对照组比较,DSS组结肠病理下可见明显损伤,DAI评分明显上升(P<0.05),炎性细胞因子IL-6、IL-1β及TRPV1、Gαq蛋白上调(P<0.05),TRPM8表达下调(P<0.05)。同时结肠黏膜及黏膜下层可见TRPV1与TRPM8、TRPV1与Gαq的共表达,TRPV1与TRPM8共表达于DRG,相较于空白对照组,DSS组结肠组织TRPV1表达增加,TRPM8表达减少。结论实验性急性结肠炎小鼠结肠组织TRPV1表达升高,TRPM8表达降低,TRPV1/TRPM8可共表达于结肠组织及DRG,两者之间可能存在某种平衡机制以维持肠道功能稳定。
Objective To investigate the expression of transient receptor potential vanilloid 1(TRPV1)and transient receptor potential melastatin member 8(TRPM8)in the intestinal and spinal dorsal root ganglion(DRG)in experimental colitis mice induced by dextran sodium sulfate(DSS),and further explore the connection and related pathways.Methods A total of 20male C57BL/6mice were selected and randomly divided into blank control group and DSS group,with 10mice in each group.Mice in the DSS group were treated with 30g/L DSS for 7days to construct colitis model.The hair,body mass,fecal characteristics(granular,loose stool,bloody stool,etc)and perianal conditions(redness,swelling,bloody stool,etc)of every group were observed and recorded at the same time every day,the colitis related disease activity index(DAI)calculated,and colon histopathological score were calculated according to the pathological sections.The protein expression levels of TRPV1,TRPM8,Gαq in colon tissue were detected by Western blot.The expression levels of interleukin-6(IL-6),interleukin-1β(IL-1β)in the colon tissue were detected by real-time quantitative polymerase chain reaction(RT-PCR).The localization and expression of TRPV1,TRPM8,Gαq in colon tissue and DRG were detected by immunofluorescence.Results Compared with the blank control group,the colon pathological damage was obvious in the DSS group,and DAI score of colon tissue was significantly increased(P<0.05),inflammatory factors including IL-1β,IL-6 and TRPV1,Gαq proteins were up-regulated significantly(P<0.05),TRPM8 expression was down-regulated(P<0.05).Meanwhile,co-expression of TRPV1/TRPM8,TR-PV1/Gαq were observed in colonic mucosa and submucosa,and TRPV1/TRPM8 were co-expression in DRG.Compared with the blank control group,TRPV1 expression was increased and TRPM8 was decreased in DSS group.Conclusion The increased TRPV1 expres sion and decreased TRPM8 expression are found in the colonic tissue of experimental colitis mice,and TRPV1/TRPM8 can be co-expressed in the colonic tissue and DRG.There may be some balance mechanism between them to maintain the stability of intestinal function.
作者
查兰兰
沈磊
吴静
余晓云
彭帅
刘佩璐
ZHA Lanlan;SHEN Lei;WU Jing(Department of Gastroenterology,Renmin Hospital of Wuhan University,Hubei Key Laboratory of Digestive System Disease,Hubei 430060,China)
出处
《医学研究杂志》
2023年第12期20-25,88,共7页
Journal of Medical Research
基金
国家自然科学基金资助项目(81570490)。
关键词
实验性结肠炎
瞬时受体电位通道
辣椒素受体
炎症性肠病
Experimental colitis
Transient receptor potential channel
Capsaicin receptor
Inflammatory bowel disease
